Team:Newcastle/14 July 2010

From 2010.igem.org

(Difference between revisions)
(→Materials)
(→Research)
 
(9 intermediate revisions not shown)
Line 1: Line 1:
{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
-
=LacI BioBrick Construction=
+
=Research=
-
==Aims==
+
We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.
-
*To use PCR to extract ''lacI'' (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
+
-
==Materials==
+
#Kleerebezem M, Bongers R, Rutten G, Vos WM de and Kuipers OP. (2004). "Autoregulation of subtilin biosynthesis in Bacillus subtilis: the role of the spa-box in subtilin-responsive promoters". ''Peptides''. 25(9). 1415-24.
-
*[[Team:Newcastle/13_July_2010|Overnight digests]]
+
#Klein C, Kaletta C, Schnell N and Entian KD. (1992). "Analysis of genes involved in biosynthesis of the lantibiotic subtilin". ''Applied and environmental microbiology''. 58(1). 132-42.  
-
*[[Team:Newcastle/8_July_2010|Ligation mix]]
+
-
*Competent ''E. Coli'' DH5alpha
+
-
 
+
-
==Protocol==
+
-
*The overnight digest are run on a gel
+
-
*Repeat transformation of DH5alpha with ligation mix
+
-
 
+
-
==Results==
+
-
The gel image was unfortunately lost, however the gel was run using all of the samples and a PCR product of known length, so as to check that the Molecular weight marker was running correctly. The results were the same as the gel run [[Team:Newcastle/13_July_2010|yesterday]], i.e. colonies 1 and 2 contain only pSB1AT3 vector (running at 3 Kbp) and the others contain pSB1AT3 with the RFP insert.
+
-
 
+
-
==Conclusion==
+
-
The conclusion that can be reached from these results is that we have not been able to get transformants with the correct plasmid/insert combination. To this end the transformation is repeated. However there is again some confusion concerning the sizes, so the samples we already have will undergo a double digest tomorrow in order to extract whatever inserts may be present.
+
{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 20:15, 25 October 2010

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

Research

We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.

  1. Kleerebezem M, Bongers R, Rutten G, Vos WM de and Kuipers OP. (2004). "Autoregulation of subtilin biosynthesis in Bacillus subtilis: the role of the spa-box in subtilin-responsive promoters". Peptides. 25(9). 1415-24.
  2. Klein C, Kaletta C, Schnell N and Entian KD. (1992). "Analysis of genes involved in biosynthesis of the lantibiotic subtilin". Applied and environmental microbiology. 58(1). 132-42.
Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
FaceBook Icon
Retrieved from "http://2010.igem.org/Team:Newcastle/14_July_2010"